藁本内酯通过降低VEGF的水平抑制人血管瘤内皮细胞的血管新生和上皮-间充质转化  

Ligustilide inhibits angiogenesis and epithelial-mesenchymal transition in human hemangioendothelial cells by reducing VEGF levels

作　　者：陈春兰[1] 张运君[1] 许和平[1] 郑少颜 CHEN Chun-lan;ZHANG Yun-jun;XU He-ping;ZHENG Shao-yan(Department of General Medicine,Hainan People's Hospital,Haikou 570100,China)

机构地区：[1]海南省人民医院全科医学科,海南海口570100

出　　处：《中国病理生理杂志》2020年第8期1422-1427,共6页Chinese Journal of Pathophysiology

基　　金：海南省自然科学基金资助项目(No.2016CXTD012)。

摘　　要：目的:探讨藁本内酯对人血管瘤内皮细胞(HemECs)的影响,并分析其机制。方法:CCK-8法检测不同浓度的藁本内酯对HemECs活力的影响;设立对照组及10、25和50μmol/L的藁本内酯处理组,EdU染色检测HemECs增殖的变化,微管形成实验检测HemECs血管新生的变化,Western blot法检测HemECs中血管内皮生长因子(VEGF)及上皮-间充质转化(EMT)相关标志物表达的变化。结果:0.1~50μmol/L的藁本内酯处理对细胞活力的影响并不明显。与对照组相比,25和50μmol/L藁本内酯处理组的Edu阳性细胞明显减少(P<0.05),微管样结构的数量明显减少(P<0.05),VEGF蛋白表达显著下调(P<0.05),E-cadherin蛋白表达显著上调(P<0.05),vimentin和β-catenin蛋白表达显著下调(P<0.05)。与对照组相比,VEGF转染组的VEGF和vimentin蛋白表达显著上调(P<0.05),E-cadherin蛋白表达显著下调(P<0.05);与VEGF转染组相比,50μmol/L藁本内酯处理VEGF转染细胞后,VEGF和vimentin蛋白表达显著下调(P<0.05),E-cadherin的蛋白表白显著上调(P<0.05)。结论:藁本内酯可抑制HemECs的增殖,还可通过降低VEGF的水平,抑制HemECs的血管新生与EMT过程。AIM:To investigate the effect of ligustilide on human hemangioendothelial cells(HemECs)and to analyze its mechanism.METHODS:The effect of ligustilide at different concentrations on the viability of HemECs was measured by CCK-8 assay. The HemECs were divided into control group and ligustilide(10,25 and 50 μmol/L)treatment groups,and the proliferation of HemECs was detected by EdU staining. The effects of ligustilide on the angiogenesis of HemECs was tested by microtubule formation experiment. The protein expression of vascular endothelial growth factor(VEGF)and epithelial-mesenchymal transition(EMT)-related markers in HemECs cells was determined by Western blot.RESULTS:There was no significant difference in the viability of the cells treated with ligustilide at the concentrations between 0. 1~50 μmol/L compared with control cells. Compared with control group,ligustilide at 25 and 50 μmol/L significantly reduced the number of EdU-positive cells and microtubule-like structures(P<0. 05),reduced the protein expression level of VEGF(P<0. 05),increased the protein expression of E-cadherin,and decreased the protein expression of vimentin and β-catenin(P<0. 05). Compared with control group,the expression of VEGF and vimentin was significantly upregulated,and the protein expression of E-cadherin was significantly down-regulated in VEGF overexpression group(P<0. 05). Compared with VEGF overexpression group,the expression of VEGF and vimentin in 50 μmol/L ligustilide-treated VEGF-overexpressing cells were significantly reduced(P<0. 05),and the protein expression of E-cadherin was significantly increased(P<0. 05).CONCLUSION:Ligulide inhibits the proliferation of HemECs,and also inhibits the angiogenesis and EMT process of HemECs by reducing the level of VEGF.

关 键 词：藁本内酯 人血管瘤内皮细胞 血管内皮生长因子 细胞增殖 血管新生 上皮-间充质转化 

分 类 号：R732.2[医药卫生—肿瘤] R730.23[医药卫生—临床医学]